The reconstitution of high 0, evolution in CaCl,-treated PS II particles was achieved by the simultaneous addition of the 17-, 23-and 34kDa proteins and total thylakoid lipids in the presence of 25% glycerol and 15 mM sodium cholate. The activity of the reconstituted membranes recovered to 85% of that of the nondepleted original PS II particles at the optimal condition. By means of this reconstitution method, evidence for the cooperation of the three proteins in the recovery of 0, evolution in the CaCl,-treated PS II particles was found by changing the concentration of NaCl in the assay medium, and the relationship between the amount of manganese retained in the water-splitting complex and the 0, evolving activity was examined by using the partially solubilized PS II particles with n-octyl-/I-D-glucoside.
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